JP2013511990A5

JP2013511990A5 -

Info

Publication number: JP2013511990A5
Application number: JP2012541211A
Authority: JP
Filing date: 2010-11-25
Publication date: 2014-01-09

Claims

A double-stranded nucleic acid molecule having the structure (A) described below,
(A) 5 ′ N ^1- (N) x-Z 3 ′ (antisense strand)
^{3 'Z'-N 2 - (} N') y-z '' 5 '( sense strand)
Wherein each of N ² , N, and N ′ is an unmodified or modified ribonucleotide, or an unconventional moiety;
Each of (N) x and (N ′) y is an oligonucleotide in which each successive N or N ′ is linked to an adjacent N or N ′ by a covalent bond;
each of x and y is independently an integer from 17 to 39;
Wherein (N ') y sequence of, it is complementary to the sequence of (N) x, wherein (N) x sequence of, it is complementary to contiguous sequences of the target RNA,
N ¹ is covalently bonded to (N) x to form Watson-Crick base pairing with N ² ,
N ¹ is selected from the group consisting of uridine, deoxyribouridine, ribothymidine, deoxyribothymidine, adenosine, or deoxyadenosine, which is mismatched and natural or modified with the target RNA , or is a non-conventional moiety,
Where z ″ may or may not be present, but if present, is a capping moiety that is covalently bonded to the 5 ′ end of N ^2- (N ′) y;
Each of Z and Z ′ is independently present or absent, but if present is independently 1 to 5 consecutive bonds covalently bound to the 3 ′ end of the chain in which it is present. A double-stranded nucleic acid molecule , or a pharmaceutically acceptable salt of said double- stranded nucleic acid, which is a nucleotide, a continuous non-nucleotide moiety, or a combination thereof .

x = y = 18, x = y = 19 or x = y = Ri 20 der, and preferably x = y = 18, the double-stranded nucleic acid molecule of claim 1.

The double-stranded nucleic acid molecule according to claim 1 , wherein N ¹ is selected from the group consisting of natural or modified adenosine and deoxyadenosine , and the corresponding nucleotide of the target RNA is adenosine , cytidine, or guanosine. .

N ¹ was natural or modified, are selected from the group consisting of uridine or deoxyuridine, the corresponding nucleotide of the target RNA, uridine, cytidine, or guanosine, double-stranded nucleic acid molecule of claim 1 .

A double-stranded nucleic acid molecule according to any one of the preceding claims , wherein Z "is present.

The double-stranded nucleic acid molecule according to any one of claims 1 to 5 , wherein Z and Z 'are absent.

The double-stranded nucleic acid molecule according to any one of claims 1 to 5 , wherein one of Z or Z 'is present.

The double-stranded nucleic acid molecule according to any one of claims 1 to 5 , wherein Z or Z 'comprises a non-nucleotide moiety.

N, N ', or at least one of N ² includes 2'OMe sugar modified ribonucleotides, a double-stranded nucleic acid molecule according to any one of the preceding claims.

The double-stranded nucleic acid molecule according to any one of the preceding claims , wherein the double-stranded nucleic acid molecule is siRNA, siNA, or miRNA.

x = y = 18, Z ′ is absent, Z is present and contains two alkyl moieties covalently bonded to each other by a phosphodiester bond;
N ² includes adenosine, or N ¹ includes natural or modified uridine, deoxyribonucleotides uridine, ribothymidine or deoxyribothymidine,
N ² includes natural or modified uridine or deoxyuridine, N ¹ includes natural or modified adenosine or deoxyadenosine ,
The double-stranded nucleic acid molecule according to claim 1.

The double-stranded nucleic acid molecule according to any one of the preceding claims , wherein the target RNA is a mammalian gene mRNA , preferably a human gene mRNA .

The double-stranded nucleic acid molecule according to any one of claims 1 to 12 , which is used in therapy.

A pharmaceutical composition comprising at least one double-stranded nucleic acid molecule according to any one of claims 1 to 12 and a pharmaceutically acceptable carrier.

Use according to any one of claims 1-12 for use in treating or preventing the occurrence or severity of a disease or condition in a subject in need of treatment or prevention of the occurrence or severity of the disease or condition. 15. A single-stranded nucleic acid molecule or a pharmaceutical composition according to claim 14, wherein the disease or condition and / or symptoms associated therewith are hearing loss, acute renal failure (ARF), organ after kidney transplantation Dysfunction (DGF), glaucoma, ocular ischemia (non-arteritic ischemic optic neuropathy (NAION), anterior ischemic optic neuropathy, age-related macular degeneration (AMD), ischemic optic neuropathy (ION) and Dry eye syndrome), acute respiratory distress syndrome (ARDS) and other acute lung and respiratory injuries, chronic obstructive pulmonary disease (COPD), primary transplant failure, ischemia reperfusion injury, reperfusion injury Primary transplantation after reperfusion edema, allograft dysfunction, lung reimplantation response and / or organ transplantation, especially after transplantation of lung transplants, organ transplants (including lung, liver, heart, pancreas, and kidney transplants) A double-stranded nucleic acid molecule selected from the group consisting of dysfunction (PGD), nephrotoxicity and neurotoxicity, spinal cord injury, brain injury, neurodegenerative disease or condition, pressure ulcer, oral mucositis, fibrotic disease, and cancer or Pharmaceutical composition .

A method of generating a double stranded RNA molecule consisting of a sense strand and an antisense strand, comprising:
a) selecting a contiguous 17-25 nucleotide sequence of the target RNA and synthesizing an antisense strand comprising complementarity to the contiguous 17-25 nucleotide sequence of the target mRNA, The 5 ′ terminal nucleotide of the antisense strand is replaced with uridine, modified uridine, ribothymidine, deoxyribothymidine, adenosine, modified adenosine, deoxyadenosine, or modified deoxyadenosine, provided that the rG: rU chain oligonucleotide and the target a step of 3 ′ terminal nucleotide of mRNA,
b) synthesizing a 17-25 nucleotide sense strand having complementarity to the antisense strand, wherein the 3 'terminal nucleotide of the sense strand and the 5' terminal nucleotide of the antisense strand Forming a Watson-Crick base pair of, and
c) annealing the sense and antisense strands, thereby producing a double stranded RNA molecule.

A method of producing a modified siRNA duplex consisting of a sense strand and an antisense strand that exhibits enhanced RNAi activity compared to an unmodified siRNA duplex comprising:
a) selecting a contiguous 17-25 nucleotide sequence of the target mRNA and synthesizing a sense strand comprising the contiguous 17-25 nucleotide sequence of the target mRNA, wherein the 3 'terminal nucleotide is Substituted with adenosine, modified adenosine, deoxyadenosine, or modified deoxyadenosine; and
b) synthesizing an antisense strand of 17-25 nucleotides complementary to the sense strand, wherein the 5 ′ terminal nucleotide is ribouridine, modified ribouridine, deoxyribouridine, or modified deoxyribouridine; and Including base pairing with the 3 'terminal nucleotide of the passenger strand;
c) annealing the sense strand to the antisense strand;
Thereby generating siRNA duplexes with enhanced RNAi activity.